BD Pathway bioimaging systems
Features
Powerful and Flexible Analysis Tools
The BD Pathway system provides analysis and visualization tools for high-content image data. This data is analyzed using visualization tools such as bar charts, scatter plots, dose-response curves, heatmaps, cell-by-cell or well-by-well analysis, cell scoring (percentage of cells responding), and z'-factor. Both endpoint and kinetic data can be analyzed using the BD Pathway system.
Visualization of high-content analysis
Using the BD Pathway™ system, bioresearchers collect and view data to gain insights into results at the cellular and subcellular level. BD Pathway software provides powerful image preprocessing filters for segmentation, including background correction, morphological, and convolution filters. These filters improve the extraction and identification of cells or cellular regions of interest for improved image analysis. The software also enables researchers to quickly and accurately split objects using sophisticated algorithms to optimize data analysis.
Phagosomal Escape Assay to Quantify Bacterial Count
Confocal collapsed stack images (40x, 0.9 NA) were taken of macrophages infected with wild-type (top) and LLO-minus mutant L. monocytogenes (bottom). The images were pseudocolored, merged, and cropped. Macrophage actin is green, bacteria are red, and colocalized signals appear as merges of these color channels. Images on right show segmentation masks generated after Sub Object counting from images on the left. The upper right plot shows time-dependent colocalization coefficients of wild-type and LLO-minus mutant bacteria with actin and vice versa, respectively (n = 4 wells). The lower right plot shows time-dependent quantification of bacterial count per macrophage (n = 4 wells).
Additional cellular and subcellular features
Intracellular imaging and analysis evaluating the cell nucleus are supported by band segmentation output, including erosion of the nuclear region. This output enables the BD Pathway system to deliver better results for plasma membrane translocation and additional subcellular region analysis. Exclusion regions can be created around the perinuclear region for improved signal to noise in other nucleus/cytoplasm translocation assays. Morphometric, intensity, and positional measurements support more precise characterization and analysis of cellular and subcellular regions or phenotypes. The BD Pathway software also counts objects within objects, which is a useful tool for applications such as DNA damage, colocalization, and spot counting.
Phagocytosis Assay
Single field confocal collapsed stack images (40x, 0.9 NA) of (A) uninfected macrophages and (B) macrophages infected with L. monocytogenes at an MOI of 128. The images were pseudocolored and merged. Macrophages are blue, bound bacteria are green, total bacteria are red, and colocalized signals appear as merges of these color channels. The log scale plot shows the effect of increasing MOI on bound and total bacterial count (number of bacteria per macrophage) in red, and total and average bacterial area per macrophage from replicate wells (n = 12 wells) in black. The bar chart shows the effect of activation of macrophages by IFN-g plus LPS on the average bound and total bacteria per macrophage (n = 4 wells).
Class I (1) laser product.
